A low-voltage alternant direct current electroporation chip for ultrafast releasing the genome DNA of Helicobacter pylori bacterium.

Abstract

Nucleic acid detection, as an important molecular diagnostic method, is widely used in bacterial identification, disease diagnosis. For detecting the nucleic acid of bacteria, the prerequisite is to release nucleic acids inside the bacteria. The common means to release nucleic acids is thechemical method, which involves complex processes, is time-consuming, and remains chemical inhibitors. Compared with chemical methods, electroporation as a physical method has the advantages of easy operation, short-time consumption, and chemical reagents free. However, the current works using electroporation often necessitates high-frequency or high-voltage conditions, entailing bulky power devices. Herein, we propose a low-voltage alternant direct current (LADC) electroporation chip and the corresponding miniature device for ultrafast releasing thegenome DNA from Helicobacter pylori (H. pylori)for detection. We connected a micrometer-interdigital electrode in the chip with a 20V portable battery to make the miniature device. Using this low-voltage device, our chip released genome DNA of H. pylori within only 5ms, achieving a cell lysis rate of 99.5%. We further combined this chip with acolorimetric loop-mediated isothermal amplification assay to visually detect H. pylori within ~ 25min at 10CFU/μL. We detected 11 clinical samples using the chip, and the detection results were consistent with those of the clinical standard. The results indicate that the LADC electroporation chip is useful for ultrafast release of genome DNA from bacteria and is expected to promote the development of nucleic acid detection in POCT and other scenarios.